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Difference in suitable mechanical properties of three‐dimensional, synthetic scaffolds for self‐renewing mouse embryonic stem cells of different genetic backgrounds
Author(s) -
Lee Myungook,
Ahn Jong Il,
Ahn Ji Yeon,
Yang Woo Sub,
Hubbell Jeffrey A.,
Lim Jeong Mook,
Lee Seung Tae
Publication year - 2017
Publication title -
journal of biomedical materials research part b: applied biomaterials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.665
H-Index - 108
eISSN - 1552-4981
pISSN - 1552-4973
DOI - 10.1002/jbm.b.33763
Subject(s) - embryonic stem cell , self healing hydrogels , microbiology and biotechnology , peg ratio , cell culture , chemistry , stem cell , polyethylene glycol , tissue engineering , gene expression , biology , gene , biochemistry , genetics , polymer chemistry , finance , economics
We evaluated whether the genetic background of embryonic stem cells (ESCs) affects the properties suitable for three‐dimensional (3D) synthetic scaffolds for cell self‐renewal. Inbred R1 and hybrid B6D2F1 mouse ESC lines were cultured for 7 days in hydrogel scaffolds with different properties derived from conjugating 7.5, 10, 12.5, or 15% (wt/vol) vinylsulfone‐functionalized three‐, four‐, or eight‐arm polyethylene glycol (PEG) with dicysteine‐containing crosslinkers with an intervening matrix metalloproteinase‐specific cleavage sites. Cell proliferation and expression of self‐renewal‐related genes and proteins by ESCs cultured in feeder‐free or containing 2D culture plate or 3D hydrogel were monitored. As a preliminary experiment, the E14 ESC‐customized synthetic 3D microenvironment did not maintain self‐renewal of either the R1 or B6D2F1 ESCs. The best R1 cell proliferation (10.04 vs. 0.16–4.39; p < 0.0001) was observed in the four‐arm 7.5% PEG‐based hydrogels than those with other properties, whereas the F1 ESCs showed better proliferation when they were embedded in the three‐arm 10% hydrogels. Self‐renewal‐related gene and protein expression by ESCs after feeder‐free 3D culture was generally maintained compared with the feeder‐containing 2D culture, but expression patterns and quantities differed. However, the feeder‐free 3D culture yielded better expression than the feeder‐free 2D culture. In conclusion, genetic background determined the suitability of hydrogel scaffolds for self‐renewal of ESCs, which requires customization for the mechanical properties of each cell line. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2261–2268, 2017.